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• O.1 mM Non-Essential Amino Acids (NEAA)
• fetal bovine serum to a final concentration of 10%

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels.
6. Incubate cultures at 37°C.

Sugimoto T, et al. Determination of cell surface membrane antigens common to both human neuroblastoma and leukemia-lymphoma cell lines by a panel of 38 monoclonal antibodies. J. Natl. Cancer Inst. 73: 51-57, 1984. PubMed: 6610792

Iavarone A, et al. Uptake and storage of m-iodobenzylguanidine are frequent neuronal functions of human neuroblastoma cell lines. Cancer Res. 53: 304-309, 1993. PubMed: 8417824

Goillot E, et al. Tumor necrosis factor as an autocrine growth factor for neuroblastoma. Cancer Res. 52: 3194-3200, 1992. PubMed: 1317260